Princess Celestia- Twilight is the student of Celestia, who is responsible for her relocation to Ponyville. They seem to share a familial bond, and Twilight has great respect for the Princess.

Rarity- Twilight has been on the receiving end of Rarity's makeovers multiple times. Rarity is one of three participants in Twilight's first sleepover. Twilight admires Rarity for her generosity and Rarity in turn admires Twilight's intellect and skills; both being unicorns, they have shared their magical skills with one another, as shown in the events.

Fluttershy - Twilight is extremely confident in Fluttershy's natural empathy with animals, even more so than Fluttershy herself.

Rainbow Dash - Twilight has been shown to be impressed by Rainbow Dash's cloud-clearing abilities. She considers Rainbow to be a brave pony.

Applejack - Twilight has been seen to appreciate Applejack's hard work, offering to help her with her farm on more than one occasion.

Pinkie Pie - Twilight enjoys the presence (or at least humors) of Pinkie Pie, but is usually shown not to have nearly the requisite energy to deal with her on a regular basis. At one point she is seen reading a book and pretending to agree with everything Pinkie Pie says in a deadpan tone.

Spike - Spike is Twilight's assistant and closest friend, having been hatched when Twilight was young as her test to get into the school for gifted unicorns. They share a lot of friendly banter and spend a lot of their time together, usually with Spike riding on Twilight's back. Spike also serves as a channel of communication between Twilight and Princess Celestia via scrolls, which he often reads as well.

Twilight Sparkle - Applejack and Twilight both respect politeness and honesty and so are very close, both valuing the skills and talents of the other. Applejack often takes the role of 'second in command' on any expeditions, directly consulting with Twilight and using her leadership abilities to motivate the others.

Fluttershy – Applejack has the most amount of patience with Fluttershy, pulling her all the way up the cliff in Dragonshy and defending her from the remarks of others.

Rainbow Dash - Applejack is good friends with Rainbow Dash, though it doesn't always seem that way. They both possess great athletic ability and love to test themselves against each other, but Rainbow Dash's rude and argumentative personality grates with Applejack's and often leads them to fight. They seem to treat this as another part of the competition however, and neither holds any grudges. Rainbow Dash is her best friend.

Pinkie Pie – Applejack's easygoing personality leads her to accept Pinkie without comment, and she states that like many residents of Ponyville she has grown used to her odd behavior. The two share an interest in baking and so get along well, with Applejack occasionally helping Pinkie out at her job.

Rarity – Uniquely among the central cast, Applejack and Rarity actively dislike each other. Applejack considers Rarity to be stuck up, condescending and finicky, while Rarity considers Applejack to be ignorant, dirty and lacking in culture. Since they share friends, both prefer to avoid each other rather than argue, though being stuck together in Look Before You Sleep allowed them to overcome some of their issues and form a somewhat better relationship.